1) 37 strains of C. tetani were isolated from 100 soil specimens collected on the surface of the ground of various places. Tetanus toxin was demonstrated in another 14 samples. Ten strains of C. tetaniwere isolated from 1 mg of earth. Tetanus toxin was demonstrated in 0.1 mg of an earth sample collected at a place. The patient who was injured at this place developed tetanus after his wound was cleaned and sutured by a surgeon.
2) C. tetani was isolated from all 10 samples of earth collected at this place and 50 cm apart one after another 2 years and 7 months later.
3) C. tetani does not remain at the same place in the same quantity. C. tetani can be isolated more readily from the soil samples collected on the surface of than deep in the ground.

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Vancomycin hydrochloride, a nonabsorbable antibiotics, which is effective against anaerobic but ineffective against aerobic gram negative rods, was administered to five consecutive cirrhotics with intractable portal-systemic encephalopathy (PSE) who were resistant to oral lactulose administration, and changes in clinical features, blood ammonia levels, electroencephalograms, and in faecal bacterial flora were examined. Vancomycin was also administered to one cirrhotic case in which administration of lactulose caused the severe diarrhoe during the use of the drug. Clinical features of encephalopathy disappeared in all cases, and electroencephalograms showed marked improvements. Faecal bacterial investigation showed that the anaerobic gram-negative rods (chiefly Bacteroides spp) were 10^8-9 counts per gram of stool on administration of lactulose, but remarkably decreased to 10^3-6 counts after administration of vancomycin. It is concluded that vancomycin is effective to the intractable PSE because of the marked decrease in anaerobic gram-negative rods.

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From April 1980 till December 1984, 2213 patients with complaint of diarrhea were bacteriologically studied in our hospital.
Examination of feces proved enteropathogenic bacteria in 612 patients. Campylobacter fetus was merely found in 12 cases (0.5%) among them, while Campylobacter jejuni and coli were detected in 418 (18.9%) and 3 (0.1%) patients, respectively.
Ten of twelve patients with Campylobacter fetus showed no clinical sign other than enteritis, however the other two had serious symptoms of systemic infection. One of these two patients had bacteremia accompanied with chill and high fever besides diarrhea: Campylobacter fetus was proved by the culture of both blood and feces. The other patient was a typical case of mother-to-baby transmission: namely, her baby had been infected with the same pathogene from her through the placenta before birth and had revealed severe meningitis and enterogastritis already at birth. Campylobacter fetus was detected from soft bloody feces of the baby as well as from the feces of the mother before and after her delivery.
The authors keenly feel the necessity of examination of feces of diarrheal patients not only for Campylobacter jejuni but also for Campylobacter fetus.

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Several biotyping schemes of Campylobacter jejuni have been proposed. A scheme of Skirrow andBenjamin (J. Clin. Pathol., 33, 1122, 1980) is based on hippurate hydrolysis and rapid H₂S test. Hebert (J. Clin. Microbiol., 15 1065, 1982) has proposed a scheme based on hippurate hydrolysis, DNA hydrolysis and growth on charcoal yeast extract agar. Recently, Lior (J. Clin. Microbiol., 20, 636, 1984) has proposed a new scheme using improved media and methods for the detection of hippurate hydrolysis, rapid H₂S production and DNA hydrolysis.
Following Lior's scheme, attempts were made to do biotyping 307 strains of Campylobacter jejuniisolated from diarrheal patients in our hospital. As reported by Lior, four biotypes were identified, 178 strains (58.0%) belonged to biotype I, 122 strains (39.7%) to biotype II, 3 strains (1.0%) to biotype III and 4 strains (1.3%) to biotype IV.
It is concluded that the Lior's biotyping scheme is easy to carry out and is useful for epidemiological study of Campylobacter jejuni infections.

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We studied the enhancement of virulence in mixed infections with E. coli and ^{K. pneumoniae}, or P.aeruginosa in mice by systemic infection of organism at a dose below the minimal lethal dose. The causes of enhanced virulence were examined for the superoxide dismutase of infecting organisms and the phagocytosis of organisms by using mouse neutrophils.
The enhanced virulence was observed in mixed infections of E. coli combined K. pneumoniae or P.aeruginosa that had high superoxide dismutase activity. i.e. the mixed infection of these oragnisms resulted in the mortality higher than those of mixed infection with low superoxide dismutase activity organisms and mono-infection by these oragnism alone. In vitro phagocytosis and killing of these organisms by mouse neutrophils, the high superoxide dismutase activity organisms are more resistant to killing effect than mono-inoculation of organism alone. It has been suggested that the enhancement of virulence in mixed infection is related to the levels of SOD in the organisms.

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Recently, we have experienced four cases of zoonosis caused one case by digs bite and three cases by cat bite and scratch. Biochemical identification and sensitivity tests were carried out on those pathogens. Pathogenic microorganisms were successfully cultivated on the blood agar but failed to growon the MacConkey's and BTB agar. All of the microorganisms showed following characteristics on agar such as positive oxidase and catalase reaction, positive indole production and glucose fermentation and also showed positive hemolysis on sheep blood agar, negative motility, and sensitive to penicillin, and the microorganism was identified as Pasteurella multocida (P. multocids). All of the antibiotics tested here were strongly effective on those isolates.
Cultures of the oral swabs taken from the dogs and cats revealed positive culture for P. multocida in 21.1%(8 of 38) and 71.4%(15 of 21), respectively.
Because infections caused by the animal bite and/or scratch were susally treated by antibiotics without microbiological examination, latent P. multocida infection as zoonosis are considered to be much more common than we have thought.
We must make more attention on the P. multocida infection, since pet keepers are increasing and severe zoonosis by animal bite and/or scratch is thought to be increase gradually.

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Minimum inhibitory concentrations (MICs) of norfloxacin (NFLX) against 100 strains of Shigella, each 25 strains of Escherichia coli, Salmonella, Campylobacter jejuni, and Vibrio (10 strains of V.parahaemolyticus, 10 strains of V. cholerae and 5 other Vibrio species), which had been isolated from enteritis patients, were measured by agar dilution method and compared with those of NA, PPA, ENX, OFLX, CPFX, FOM and KM. The MICs of NFLX which inhibited 90% of the strains were 0.10μg/ml against Shigella and Salmonella, 0.39μg/ml against E. coli, 0.20μg/ml against V parahaemolyticus and other Vibrio strains, 0.05μg/ml against V cholerae, and 1.5μg/ml against C. jejuni. NFLX inhibited all of the tested strains at the concentration of 3.13μg/ml or below. These values were substantially lower than those of NA, PPA, FOM and KM and roughly equal to those of ENX, OFLX and CPFX.
Three NA-resistant Shigella strains were all sensitive to NFLX.

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A medical staff of Osaka University Hospital (31-year-old male) was admitted to our department because of hemorrhagic fever with renal syndrome (HFRS) in November, 1984. He only visited an animal laboratory two weeks before the onset of fever but touched neither animalsnor their discharges. His laboratory data were first obtained on the day of onset of fever and followedalmost daily throughout one-month hospitalization. The clinical examinations demonstrated the deveropmentof disseminated intravascular coagulation syndrome (DIC), hepatic, renal and cardiac damages withelectrolyte and metabolic disorders. This is the first HFRS case in whom urinary β₂-microglobulin and N-acetyl-β, Dglucosaminidase, isoenzyme patterns of serum LDH and CPK were examined.
Titers of serum antibody to HFRS viruses (both Hantaan 76-118 and B-1 strain) were measured by indirect fluorescent antibody (IF) and hemagglutination inhibition (HI) tests throughout the course. The titers obtained by these two methods were changed in parallel, but IF test was proved to be more sensitive and useful for earlier diagnosis of HFRS. The titer of antibody to B-1 strain measured by IF test was elevated to 1: 128 on the third day from the onset and reached maximum levels of 1: 32, 000-64, 000 after two weeks.

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For the early diagnosis of systemic candidiasis which has been increasing in recent years, we developed an enzyme-linked immunosorbent assay with avidin-biotin system (A-B ELISA) to detect serum Candida antigen. A-B ELISA was evaluated in murine systemic candidiasis.
A-B ELISA detected purified C. albicans mannan with a sensitivity of 1.2 ng/ml inpooled human sera, and 0.25 ng/ml in PBS. Except for C. tropicalis C. guilliermondii and C. stellatoidea, no cross reactivities were seen with a variety of other fungal and bacterial antigen preparations. Anti C. albicans antibody which was used in this method was shown a specific binding to purified Candida mannan by ELISA inhibition technique.
In experimental systemic candidiasis of mice, serum Candida antigen was positive in 14 of 26 mice, but blood culutre was negative except for one. There was a relationship between serum antigen concentration and an area of abscesses in renal cortex and total organs of infected mouse. On the other hand, even if the foci had developed in large area, Candida antigen was not detected when the lesion had been transformed from abscess to granuloma. Therefore it is important for the diagnosis of systemic candidasis when the sample should be taken.
These results suggest that A-B ELISA to detect Candida mannan antigen is an useful method for diagnosis of systemic candidiasis because of its simple and timesaving feature and high sensitivity.

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The clinical efficacy and safety of MK-0787/MK-0791 were compared with piperacillin (PIPC) in respiratory tract infections by a well controlled study. Drugs were administered by intravenous drip infusion twice a day for 14 days, at a daily of 1 g/1 g of MK-0787/MK-0791 or 4 g of PIPC.
The following results were obtained.
1. Out of 367 patients (183 administered MK-0787/MK-0791, 184 PIPC) included in this trial, the clinical efficacy of 301 patients (155 received MK-0787/MK-0791, 146 PIPC) was evaluated by the committee members.
2. The clinical efficacy rates in all cases evaluated by the committee members were 80.0% in MK-0787/MK-0791 and 71.7% in PIPC, respectively, and no significant difference was observed between the two drugs. Wheras, the clinical efficacy rates evaluated by the attending physicians were 82.0% in MK-07871/MK-0791 and 72.0% in PIPC, and significant difference was observed between the two drugs.
3. No significant difference was observed between the clinical efficacy of the two drugs evaluated by the committee members in patients with pneumonia or pulmonary suppuration. But in patients with chronic respiratory tract infections, the clinical efficacy rates were 83.5% in MK-0787/MK-0791 and 63.0% in PIPC, respectively, therefore MK-0787/MK-0791 was significantly superior to PIPC. The clinical efficacy evaluated by both the attending physicians and the committee members in those patients were observed to be similar.
4. The clinical and bacteriological efficacy against causative organisms in all cases were observed to have no significant difference between the two drugs, but MK-0787/MK-0791 was significantly superior to PIPC in single infections by P. aeruginosa.
5. No significant difference was observed between the two drugs in regard to side effects or abnormality in laboratory findings.
6. The usefulness of MK-0787/MK-0791 was superior to PIPC in all cases and in patients with chronic respiratory tract infections judged by committee members, and in patients with chronic respiratory tract infections judged by attending physicians.
From these results it was concluded that MK-0787/MK-0791 is an extremely useful drug in the treatment of respiratory tract infections.

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Chlamydiazyme developed by Abbott laboratories, U. S. A., is an enzyme immunoassay kit to detect chlamydial antigen. The usefulness of Chlamydiazyme for the diagnosis of Chlamydiatrachomatis infection in the genitourinary tract was investigated by comparison with cell culutre method. Urethral (male) and cervical swab specimens obtained from patients attending urological, and obstetric and gynecological clinics were tested according to the manual enclosed in Chlamydiazyme kit. A net absorbance value of 1.00 or greater was determined as a positive reaction. Among 486 untreated cases (323males and 163 females), Chlamydiazyme was positive in 132 (96.4%) of 137 culture-positive specimens and negative in 322 (92.3%) of 349 culture-negative specimens. For male patients, 102 (99.0%) of 103 culture-positive specimens were Chlamydiazyme-positive and 201 (91.4%) of 220 culture-negative specimens were Chlamydiazyme-negative. In female cases the positive-coincidnece ratio (sensitivity) of the Chlamydiazyme test to cell culture was 88%(30/34) and the negative-coincidence ratio (specificity) of Chlamydiazyme was 93.8%(121/129). The positive-coincidence ratio in female cases was slightly lower than that in male cases. High net absorbances, 2.00 or greater, were observed in 50.9% of 157 positive specimens. In 62 cases which had been culture-and Chlamydiazyme-positive before treatment, 16 (73%) of 22 culture-negative specimens were Chlamydiazyme-negative during treatment, although 39 (98%) of 40 culture-positive specimens were Chlamydiazyme-positive. The low positive coincidence ratio during treatment might be due to chlamydial antigen persisting in the infected focus for one to two weeks after the onset of treatment, even if the C. trachomatis was not alive. No influence was also observed on the results of Chlamydiazyme by storage of specimens up to 4 weeks at 4°C. The Chlamydiazyme kit will provide a rapid, rather simpler than cell culture, sensitive and specific tool for the diagnosis of C. trachomatis infection in the genitourinary tract.

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The role of immunological reaction has recently been emphasized in relation to the mechanism development of pneumonia in Mycoplasma pneumoniae (M. pn.) infection, and authors analyzed lymphocyte subsets in peripheral blood in 11 patients with acute stage of M. pn. pneumonia.
In the peripheral blood lymphocyte subsets, the OKT 4/8 ratio was significantly lowered to 0.97±0.45 (p<0.001) compared with that of the healthy control group, and this was due to a decrease of the count of OKT4 positive lymphoeytes. This result may suggest inhibition of cellular immunity in the disease.

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